Li Zhanjie, Wang Kai
Key Laboratory of Genetics, Breeding and Multiple Utilization of Crops, Ministry of Education, Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, Fujian Agriculture and Forestry University, Fuzhou, China.
National Engineering Research Center of Sugarcane, Fujian Agriculture and Forestry University, Fuzhou, China.
Methods Mol Biol. 2020;2072:85-99. doi: 10.1007/978-1-4939-9865-4_8.
cis-regulatory DNA elements (CREs) are noncoding but functional DNA sequences. The binding of regulatory proteins into CRE regions leads to chromatin high sensitive to DNase I digestion, which are termed as DNase I hypersensitive sites (DHSs). These DHSs can be efficiently detected through DNase I digestion followed by high-throughput DNA sequencing (DNase-seq). Thus, DNase-seq has become a powerful technique for DHSs mapping at whole-genome level in both plants and animals. Here we describe a DNase-seq procedure modified and developed for crop plants. These plants usually contain large amounts of repetitive sequences and complex organic constituents. With the main improvement in nuclei isolation, this method has been successfully used in mapping DHSs in cotton and sugarcane.
顺式调控DNA元件（CREs）是非编码但具有功能的DNA序列。调控蛋白与CRE区域的结合会导致染色质对DNase I消化高度敏感，这些区域被称为DNase I超敏位点（DHSs）。通过DNase I消化后进行高通量DNA测序（DNase-seq），可以有效地检测到这些DHSs。因此，DNase-seq已成为在植物和动物全基因组水平上绘制DHSs的强大技术。在这里，我们描述了一种为农作物改良和开发的DNase-seq程序。这些植物通常含有大量的重复序列和复杂的有机成分。通过在细胞核分离方面的主要改进，该方法已成功用于绘制棉花和甘蔗中的DHSs。
